SIM SF Expression Medium (For SF9 and SF21, Serum free)

Product description
SIM SF Expression Medium is an optimized serum-free medium used to support the growth of SF9 and SF21 cells. It is suitable for the establishment of an efficient insect-baculovirus expression system. This medium contains L-glutamine and is ready-to-use.
Product features
• Specifically optimized for the culture and transient transfection of SF9/SF21 cells
• Strict quality control, stable performance, cost-effective
• Serum-free, protein-free
Product parameters
appearance Clear Yellow
Osmotic pressure 340-400 mOsm/kg
pH 6.0-6.5
Sterility No Microbial Growth Detected
endotoxin < 10 EU/mL
Period of validity 12 months from the date of manufacture
Storage conditions 2-8 ℃; Protect from light
* SIM SF medium is yellow transparent liquid. Do not use the medium, once it becomes cloudy or precipitated.
Culture Conditions
• Media: SIM SF Expression Medium
• Cell Line: SF9/SF21 cells
• Culture Type: Adherent or suspended culture
• Culture Vessel: Shake flask, bioreactor, etc.
• Temperature Range: 26℃ to 28℃
Recovery

1. Rapidly thaw (less than 2 minute) avail of frozen SF9/SF21cells in a 27℃ water bath;

2. Decontaminate the vial and transfer the thawed cells into a sterile centrifuge tube with 5-10 mL fresh medium;

3. Centrifuge the cells at 200 × g for 5 minutes, remove the supernatant slowly.

4. Suspend the cells with fresh medium in sterile shake flask, and culture in an orbital shaker at 150 rpm;

5. Subculture the cells when the cell density is ≥2-4× 106 viable cells/mL.

Subculture

(1). Suspended culture

1. Seed cells into pre-warmed fresh SIM SFExpression Medium at a final density of 0.8-1.0×106 viable cells/mL. Incubate the cells at 27±1℃ rotating at 150 rpm(The speed may vary among different shakers. Resuscitated cells are recommended for adaptive culture at a lower rotational speed);

2. Subculture the cells again, once cell density reaches 2-3 ×106 viable cells/mL.

(2). Adherent culture
For baculovirus packaging

1. Determine the density of cells in logarithmic phase with a viability ≥ 90% and count the cell number;

2. Inoculated 2-4×104 cells/cm2 cells into the disposable culture dish. It can be used for virus packaging after standing for more than 45minutes.

Cryopreservation

1. Determine the density of cells in logarithmic phase with a viability ≥ 90%;

2. Prepare cell freezing medium which is composed of 45% fresh SIM SF Expression Medium, 45% conditioned medium (cell culture supernatant of the cells used for cryopreservation) and 10% DMSO;

3. Centrifuge(200 × g, 5minutes) and remove all the supernatant. Resuspend the cells with suitable volume of cell freezing medium to viable cell density;

4. Aliquot the cells to cryovials and place to controlled-rate freezing apparatus. Put the apparatus in -80℃ refrigerator;

5. Transfer the tubes to liquid nitrogen for long-term storage.

Cell adaptation

(1).Direct adaptation

In general, SF9/SF21 cells can be directly adapted from serum-reduced media or other serum-free media into SIM SF Expression Medium. Cells used for adaptation is required to be in the logarithmic phase with a viability ≥ 90%.

1. Subculture the cells directly into 20 mL of fresh SIM SF Expression Medium at a final density of 1.0×106 viable cells/mL;

2. Incubate the cells at 27℃, rotating at 150 rpm;

3. 3-4 days later, determine the cell density and viability. Generally, the cell density should be >2×106 cells/mL, which means that direct adaptation is successful.

(2). Sequential adaptation

The cell inoculation density was also 1.0×106 cells/mL. In the passage process, the ratio between the primary used medium and SIM SF Expression Medium in the mixed medium gradually decreased from 90:10 to 75:25, 50:50, 25:75, 10:90, 0:100.

Note: In the process of cell adaptation, the first three generations of cells may not be good, but generally the cell state will improve from the fourth generation.

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